Research Article
Open Access Peer-reviewed

Development of a Simple HPLC Method for the Sea-lamprey Pheromone, 3-ketopetromyzonol Sulfate (3kPZS)

Kenneth Banks1, James E. Hanson1,

1Department of Chemistry and Biochemistry, Seton Hall University, South Orange NJ, USA

American Journal of Marine Science. 2021, 9(1), 12-15. DOI: 10.12691/marine-9-1-3
Received September 27, 2021; Revised October 28, 2021; Accepted November 04, 2021

Abstract

The development of a simple HPLC method for detection of the sea lamprey pheromone 3-ketopetromyzonol sulfate (3kPZS) is reported. The ketone moiety of the bile alcohol reacts readily with 1-dimethylaminonaphthalene-5-sulfonyl (dansyl) hydrazine to form the hydrazone. This hydrazone has a strong UV absorbance at 333 nm and visible fluorescence at 518 nm. Solutions containing the dansyl derivatized 3kPZS are readily analyzed by reverse phase HPLC with UV detection. A simple water-methanol gradient elutes the derivatized 3kPZS in just over 10 minutes using a standard 150 mm column. Detector response has high linearity throughout the ppm range, and the detection limit by UV is below 100 ppb. This methodology is useful for high throughput analysis of large numbers of samples in the ppm range.

Keywords:

HPLC, derivatization, bile salts, sea lamprey
[1]  Wagner, C.M., Hanson, J.E., Meckley, T.D., Johnson, N.S., Bals, J.D. A simple, cost-effective emitter for controlled release of fish pheromones: Development, testing, and application to management of the invasive sea lamprey. PLOS One, 13 (6): e0197569, 2018.View Article  PubMed
 
[2]  Li, W., Scott, A.P., Siefkes, M.J., Yan, H., Liu, Q., Yun, S.S., Gage, D.A. Bile Acid Secreted by Male Sea Lamprey that Acts as a Sex Pheromone. Science, 296, 138-141, 2002.View Article  PubMed
 
[3]  Yun, S.S., Scott, A.P., Li, W. Pheromones of the male sea lamprey, Petromyzon marinus L.: structural studies on a new compound, 3-keto allocholic acid, and 3-keto petromyzonol sulfate. Steroids, 68, 297-304, 2003.View Article
 
[4]  Johnson, N.S., Yun, S.S., Thompson, H.T., Brant, C.O., Li, W. A synthesized pheromone induces upstream movement in female sea lamprey and summons them into traps. Proc. Nat. Acad. Sci. 106 (4), 1021-1026, 2009.View Article  PubMed
 
[5]  Li, K., Scott, A.M., Chung-Davidson, Y.W., Bussy, U., Patel, T., Middleton, Z.E., Li, W. Quantification of Oxidized and Unsaturated Bile Alcohols in Sea Lamprey Tissues by Ultra-high Performance Liquid Chromatography-Tandem Mass Spectrometry. Molecules. 21 (9), 1120-1129, 2016.View Article  PubMed
 
[6]  Hagey, L.R., Moller, P.R., Hofmann, A.F., Krasowski, M.D. Diversity of Bile Salts in Fish and Amphibians: Evolution of a Complex Biochemical Pathway. Physiological and Biochemical Zoology, 83 (2), 308-321, 2010.View Article  PubMed
 
[7]  Appelblad, P., Jaegfeldt, E.P.H., Backstrom, T., Irgum, K. Derivatization of Steroids with Dansylhydrazine Using Trifluoromethanesulfonic Acid as Catalyst. Analytical Chemistry, 69 (23), 4905-4911, 1997.View Article  PubMed
 
[8]  Kawasaki, T., Maeda, M., Tsuji, A. Determination of plasma and urinary cortisol by high-performance liquid chromatography using fluorescence derivatization with dansyl hydrazine. J. Chromatography B 163 (2), 143-150, 1979.View Article
 
[9]  Chayen, R., Dvir, R., Gould, S., Harell, A. 1-Dimethyl-aminonaphthalene-5-sulfonyl hydrazine (dansyl hydrazine): a fluorometric reagent for carbonyl compounds Analytical Biochem. 42 (1), 283-286, 1971.View Article
 
[10]  The point at 100 ppm has extremely high leverage, leading to the unusually good fit. Dunn, P.K.; Smyth, G.K. “Generalized Linear Models with Examples in R” Springer, New York: 2018.
 
[11]  Xi, X.; Johnson, N.S.; Brant, C.O.; Yun, S.S.; Chambers, K.L.; Jones, A.D.; Li, W. Quantification of a male sea lamprey pheromone in tributaries of Laurentian Great Lakes by liquid chromatography-tandem mass spectrometry Environ. Sci. Technol. 45 (15), 6437-6443, 2011.View Article  PubMed
 
[12]  Wang, H.; Johnson, N.; Bernardy, J.; Hubert, T.; Li, W. Monitoring sea lamprey pheromones and their degradation using rapid stream-side extraction coupled with UPLC-MS/MS. Journal of Separation Science 36 (9-10), 1612-1620, 2013.View Article  PubMed
 
[13]  Park, S.; Jung, S.; Kim, H. Water 11, 2645-2661, 2019.View Article
 
[14]  Johnson, N.S.; Siefkes, M.J.; Wagner, C.M.; Dawson, H.; Wang, H.; Steeves, T.; Twohey, M.; Li, W. A synthesized mating pheromone component increases adult sea lamprey (Petromyzon marinus) trap capture in management scenarios Canadian Journal of Fisheries and Aquatic Sciences, 70 (7), 1101-1108, 2013.View Article
 
[15]  Brant, C.O.; Huertas, M.; Li, K.; Li, W. Mixtures of Two Bile Alcohol Sulfates Function as a Proximity Pheromone in Sea Lamprey PLoS ONE, 11 (2), e0149508, 2016.View Article  PubMed
 
[16]  Chatzimichail, S.; Rahimi, F.; Saifuddin, A.; Surman, A.J.; Taylor-Robinson, S.D.; Salehi-Reyhani, A. Hand portable HPLC with broadband spectral detection enables analysis of complex polycyclic aromatic hydrocarbon mixtures Communications Chemistry, 4, 2021.View Article